Genetic engineering of cell lines using lentiviral vectors to achieve antibody secretion following encapsulated implantation

Autor: Bernd Bohrmann, Erhard Kopetzki, Christoph Schweitzer, Patrick Aebischer, Helmut Jacobsen, Bernard L. Schneider, Aurélien Lathuilière, Marc Moniatte
Rok vydání: 2013
Předmět:
Glycosylation
Subcutaneous Fat / metabolism
Cell
Myoblasts
Transduction (genetics)
Mice
Drug Delivery Systems
Lentivirus / genetics
Transgenes
Cloning
Molecular
Promoter Regions
Genetic
biology
Immunoglobulin G / administration & dosage
Macroencapsulation
Recombinant Proteins / administration & dosage
Cells
Immobilized
Recombinant Proteins
medicine.anatomical_structure
Mechanics of Materials
Lentiviral vector
Antibody
Genetic Engineering / methods
Genetic Engineering
C2C12
Monoclonal antibody
medicine.drug_class
Transgene
Genetic Vectors
Biophysics
Subcutaneous Fat
Bioengineering
Enzyme-Linked Immunosorbent Assay
Viral vector
Cell Line
Biomaterials
medicine
Animals
Humans
Myoblasts / cytology
Passive immunization
Lentivirus
Genetic Therapy
Molecular biology
Cell encapsulation technology
Cell culture
Immunoglobulin G
Ceramics and Composites
biology.protein
Cells
Immobilized / cytology
Zdroj: Biomaterials, Vol. 35, No 2 (2014) pp. 792-802
ISSN: 1878-5905
0142-9612
Popis: The controlled delivery of antibodies by immunoisolated bioimplants containing genetically engineered cells is an attractive and safe approach for chronic treatments. To reach therapeutic antibody levels there is a need to generate renewable cell lines, which can long-term survive in macroencapsulation devices while maintaining high antibody specific productivity. Here we have developed a dual lentiviral vector strategy for the genetic engineering of cell lines compatible with macroencapsulation, using separate vectors encoding IgG light and heavy chains. We show that IgG expression level can be maximized as a function of vector dose and transgene ratio. This approach allows for the generation of stable populations of IgG-expressing C2C12 mouse myoblasts, and for the subsequent isolation of clones stably secreting high IgG levels. Moreover, we demonstrate that cell transduction using this lentiviral system leads to the production of a functional glycosylated antibody by myogenic cells. Subsequent implantation of antibody-secreting cells in a high-capacity macroencapsulation device enables continuous delivery of recombinant antibodies in the mouse subcutaneous tissue, leading to substantial levels of therapeutic IgG detectable in the plasma. (C) 2013 Elsevier Ltd. All rights reserved.
Databáze: OpenAIRE
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